Bento Ananda de Araujo, Maciel Marianna Cardoso, Bezerra Francisco Felipe, Mourão Paulo Antônio de Souza, Pavão Mauro Sérgio Gonçalves, Stelling Mariana Paranhos
Federal Institute of Education, Science and Technology of Rio de Janeiro, Rio de Janeiro 20271-110, Brazil.
Medical Biochemistry Institute, Federal University of Rio de Janeiro, Rio de Janeiro 21941-971, Brazil.
Pharmaceuticals (Basel). 2023 Oct 3;16(10):1401. doi: 10.3390/ph16101401.
Ascidians are marine invertebrates that synthesize sulfated glycosaminoglycans (GAGs) within their viscera. Ascidian GAGs are considered analogues of mammalian GAGs and possess great potential as bioactive compounds, presenting antitumoral and anticoagulant activity. Due to its worldwide occurrence and, therefore, being a suitable organism for large-scale mariculture in many marine environments, our main objectives are to study GAGs regarding composition, structure, and biological activity. We also aim to develop efficient protocols for sulfated polysaccharides extraction and purification for large-scale production and clinical applications. GAGs derived from viscera were extracted by proteolytic digestion, purified by ion-exchange liquid chromatography, and characterized by agarose gel electrophoresis and enzymatic treatments. Anticoagulant activity was evaluated by APTT assays. Antitumoral activity was assessed in an in vitro model of tumor cell culture using MTT, clonogenic, and wound healing assays, respectively. Our results show that presents three distinct polysaccharides; among them, two were identified: a dermatan sulfate and a fucosylated dermatan sulfate. Antitumoral activity was confirmed for the total polysaccharides (TP). While short-term incubation does not affect tumor cell viability at low concentrations, long-term TP incubation decreases LLC tumor cell growth/proliferation at different concentrations. In addition, TP decreased tumor cell migration at different concentrations. In conclusion, we state that presents great potential as an alternative GAG source, producing compounds with antitumoral properties at low concentrations that do not possess anticoagulant activity and do not enhance other aspects of malignancy, such as tumor cell migration. Our perspectives are to apply these molecules in future preclinical studies for cancer treatment as antitumoral agents to be combined with current treatments to potentiate therapeutic efficacy.
海鞘是一种海洋无脊椎动物，其内脏能合成硫酸化糖胺聚糖（GAGs）。海鞘GAGs被认为是哺乳动物GAGs的类似物，具有作为生物活性化合物的巨大潜力，具有抗肿瘤和抗凝血活性。由于其在全球范围内都有分布，因此是许多海洋环境中大规模海水养殖的合适生物，我们的主要目标是研究GAGs的组成、结构和生物活性。我们还旨在开发高效的硫酸化多糖提取和纯化方案，用于大规模生产和临床应用。从内脏中提取的GAGs通过蛋白水解消化法提取，通过离子交换液相色谱法纯化，并通过琼脂糖凝胶电泳和酶处理进行表征。通过活化部分凝血活酶时间（APTT）测定评估抗凝血活性。分别使用MTT、克隆形成和伤口愈合试验在肿瘤细胞培养的体外模型中评估抗肿瘤活性。我们的结果表明，存在三种不同的多糖；其中两种已被鉴定：一种硫酸皮肤素和一种岩藻糖基化硫酸皮肤素。总多糖（TP）的抗肿瘤活性得到证实。虽然短期孵育在低浓度下不影响肿瘤细胞活力，但长期TP孵育在不同浓度下会降低LLC肿瘤细胞的生长/增殖。此外，TP在不同浓度下降低了肿瘤细胞的迁移。总之，我们指出，作为一种替代的GAG来源具有巨大潜力，能产生低浓度下具有抗肿瘤特性的化合物，这些化合物不具有抗凝血活性，也不会增强恶性肿瘤的其他方面，如肿瘤细胞迁移。我们的展望是将这些分子应用于未来的癌症治疗临床前研究，作为抗肿瘤药物与当前治疗方法联合使用，以增强治疗效果。
